47
NEWGEN Zlín Workshop and WG Meeting On Hydrogel/ Biomineralized Biomaterial for Bone Tissue Regeneration 15th - 16th November 2016
NEWGEN Zlín Workshop and WG Meeting
On
Hydrogel/ Biomineralized Biomaterial for Bone
Tissue Regeneration
15th - 16th November 2016
Copyright © 2016 by Tomas Bata University in Zlín
All rights reserved
Editor: Assoc. Prof. Nabanita Saha, PhD.
Published by: Tomas Bata University in Zlín, 2016
1
Index
Serial
Number Content
Page
Number
1 MPNS COST Action MP1301 2
2 Committees 3
3 Scientific Working Group 4
4 Workshop Venue 5
5 Scientific Program – Day 1 6
6 Scientific Program – Day 2 7
7 Technical Session 8-11
8 Abstracts – Invited Speakers 12-16
9 Abstracts – Oral Speakers 17-27
10 Abstracts – Posters 28-44
2
COST Actions Materials, Physics and
Nanosciences (MPNS) MP1301 (MPNS COST Action MP1301)
New Generation Biomimetic and Customized
Implants for Bone Engineering
General Information
Chair of the Action:
Dr Francis CAMBIER (BE)
Vice Chair of the Action:
Dr Paola PALMERO (IT)
Science officer of the Action:
Dr Mónica PÉREZ-CABERO
Administrative officer of the Action:
Ms Milena STOYANOVA
3
Committees
Organizing Committee (Scientific)
Ms. Nabanita Saha, (TBU in Zlin,CPS)
Mr. Vladimír Sedlařík
Mr. Petr Humpolíček
Ms. Oyunchmeg Zaandra
Ms. Lenka Jelinková
Mr. Ramasubbareddy Palem
Mr. Ayan Ray
Mr. Stéphane Hocquet, (INISMA – CRIBC, Mons, Belgium) STSM
Coordinator
Organizing Committee (Administrative)
Ms. Nibedita Saha (TBU in Zlin,UNI/CPS), Co-PI of COST_MP1301
Ms. Monika Kasálková
Ms. Eliška Mikuličková
Ms. Nikola Mikušová
Ms. Jana Josefiková
Mr. Jiří Jaroš
Ms. Lucie Romanová
Ms. Véronique Huart (INISMA – CRIBC, Mons, Belgium) Grant Holder
Manager
4
Scientific Working Group
Working Group 1 (WG-1)
Hydrogel Scaffolds / Biomaterials
Working Group 2 (WG-2)
Bone Tissue Regeneration / Engineering
Working Group 3 (WG-3)
Drug Delivery
Working Group 4 (WG-4)
In vitro and in vivo tests
5
Workshop Venue
Address
Centre of Polymer Systems
Trida Tomase Bati 5678
760 01 Zlín
http://newgen.utb.cz/
6
Hydrogel/Biomineralized Biomaterial for Bone Tissue Regeneration
Scientific Program
The preliminary schedule for MP-1301-Workshop and WG meeting:
1st Day - 15 November 2016, Tuesday
08.30-09.00: Registration of the participants in the Workshop
09.00-09.05: Welcome from Rector, TBU in Zlin (Prof. Petr Saha)
09.05-09.30: Welcome from the MP-1301_COST Action coordinator (Dr. Francis Cambier)
09.30-10.00: Invited Talk (Assoc. Prof. Radostina Alexandrova, Bulgaria)
10.00-10.30: Coffee break and POSTER Session
10.30-11.00: Invited Talk (Prof.Yannis Missirlis, Greece)
11.00-12.00: Technical Session
11.00-11.20: David C Bassett (Norway)
11.20-11.40: Hai jun Xiao (Czech Republic)
11.40-12.00: Smarak Bandopadhyay (Czech Republic)
12.00-12.15: NEWGEN meeting in Greece in 2017 (Prof.Yannis Missirlis)
12.30-13.30: Lunch
13.30-14.00: CPS laboratory visit
14.00-14.30: Planery Talk (Prof. Jaroslav Cihlar, Brno, Czech Republic)
14.00-15.00: Technical Session
14.00-14.20: Karine Salim (France)
14.20-14.40: Antonio Di Martino (Czech Republic)
14.40-15.00: Probal Basu (Czech Republic)
15.00-15:40: Coffee break and POSTER Session [Technical Poster Session]
15.40-16.20: Technical Session
15.40-16-00: Bojana Obradovic (Serbia)
16.00-16-20: Alena Pavelkova(Czech Republic)
16.20-17.00: Working Group meeting (WG-1,WG-2, WG-3and WG-4)
17.00-19.00: end of 1st day [Resting period]
19.00h: Workshop Dinner
7
Hydrogel/Biomineralized Biomaterial for Bone Tissue Regeneration
Scientific Program
The preliminary schedule for MP-1301-Workshop and WG meeting:
2nd Day – 16 November 2016, Wednesday
08.30-09.00 : Registration of the participants in the Workshop
09.00- 09.30: Invited Talk (Prof. Martijn van Grienven, Germany)
09.30-10.00: Invited Talk (Dr. Stephe Scheiner, Austria)
10.00-10.30: Coffee break and POSTER Session
10.30-11.30: Technical Session
10.30-10.50: Silvia Ramirez (France)
10.50-11.10: Filatova Katerina (Czech Republic)
11.10-11.30: Ayan Ray (Czech Republic)
11.30-11.45: NEWGEN meeting in Vienna in 2017 (Dr. Stephan Scheiner)
11.45-12.00: Vote of Thanks (Assoc. Prof. Vladimír Sedlařík)
12.30-13.30: Lunch
13.30-15:30:Visit of Bata Museum and Skyscraper (21) Building
15.30h: end of 2nd day
8
Technical Session
Serial
Number Authors Name Title
Page
Number
1 Radostina
Alexandrova
Searching for new materials for bone implants:
challenges and solutions of in vitro biocompatibility
assessment
Radostina Alexandrova, Tanya Zhivkova, Lora Dyakova,
Boyka-Andonova-Lilova, Abedulkadir Abudalleh, Milena
Georgieva, Georgi Miloshev, Nabanita Saha, Diana
Rabadjieva, Stefka Tepavitcharova
12
2 Yannis
F.Missirlis Biomaterials’ Elicited Messages Influence Epigenetics
Yannis F.Missirlis 13
3 Jaroslav Cihlar
Structurally And Functionally Graded Biocomposites
And Scaffolds Based On Ca-Phosphates For Bone Tissue
Engineering
Cihlar J., Drdlik D., Novotna L.,Castkova K., Cihlar J. jr,
14
4 Martijn van Griensven
(Pre-)Clinical Bone Engineering Also Taking Co-
Morbidities Into Account
Martijn van Griensven
15
5 Stefan Scheiner
Stiffness And Strength Prediction For A
Hydroxyapatite-Based Biomaterial, Considering Bone
Regeneration
Stefan Scheiner, Vladimir R. Komlev, Alexey N. Gurin,
Christian Hellmich
16
6 Bassett David
Control of gelling kinetics in ionotropic polymers
applied to 3D cell friendly encapsulation
David C Bassett, Armend G. Håti, Thor B Melø, Bjørn T.
Stokke, Pawel Sikorski
17
7 Haijun Xiao Effects of X-shaped reduction-sensitive amphiphilic
block copolymer on drug delivery
Haijun Xiao, Lu Wang
18
8 Smarak
Bandyopadhyay
“Bacterial Cellulose” from Apple Juice Medium: A
Sustainable Raw Material for Hydrogel
Smarak Bandyopadhyay, Zandraa Oyunchimeg, Lenka
Jlinkova, Nabanita Saha and Petr Saha
19
9 Karine Salim Synthetic copolymers for biomedical needs:
from resorbable to permanent biomaterials
Karine Salim, Eric.Busseron, Didier Combis
20
9
Serial
Number Authors Name Title
Page
Number
10 Antonio
Di Martino Chitosan : A versatile tool for drug and gene delivery
Antonio Di Martino 21
11 Probal Basu Functional Significance of Three Dimensional Hydrogel
Scaffolds in Bone Tissue Engineering – A Review
Probal Basu, Nabanita Saha, Petr Saha
22
12 Obradovic
Bojana
Utilization of biomimetic bioreactors for biomaterial
evaluation: gellan-gum hydrogels with nano-particulate
bioactive-glass Zvicer J., Gantar A., Kocen R., Veljovic Dj., Novak S.,
Obradovic B.
23
13 Alena
Pavelková1
Non-toxic biodegradable polyester urethanes based on
PLA Alena Pavelková1, Pavel Kucharczyk, Zdenka Kuceková
Jiří Zedník and Vladimír Sedlařík
24
14 Silvia Ramirez
Mineralization process of chitosan hydrogels by calcium
phosphate apatite
S. Ramirez, L. David, T. Delair, E. Maire, A. Montembault,
S. Tadier, L. Gremillard
25
15 Filatova
Katerina
Polymer-coated silica particles for controlled drug
delivery
Filatova Katerina, Vladimir Sedlarik
26
16 Ayan Ray Biomineralized Magnetic Hydrogel: An Application
Towards Biomedicine
Ayan Ray, Nabanita Saha, Tomas Saha and Petr Saha
27
17 Marina
Sokolova
Hyaluonan hydrogel/calcium phosphates composites for
medical application
Marina Sokolova, Janis Locs, Dagnija Loca, Ilze Balode
28
18 Lucie
Nogolova
The Effect of Antimicrobial Agents on the Stability of
Hydrogels
Lucie Nogolova, Vladimir Sedlarik 29
19 Sylwia
Dąbrowska
The degradation tests of ceramic-polymer composites
with hydroxyapatite for orthopedic implants
Sylwia Dąbrowska, Elżbieta Pietrzykowska,
Roman Mukhovskyi, Agnieszka Chodara,
Tadeusz Chudoba, Janis Locs, Witold Łojkowski
30
10
Serial
Number Authors Name Title
Page
Number
20 Novotna Lenka. Mechanical and biological evaluation of calcium
phosphate / silica based macroporous scaffolds
Novotna L., Chlup Z., Cihlar J.
31
21 Maja Dutour
Sikirić
Development of injectable bone susbstitute materials
based on calcium phosphates and reverse
thermoresponsive polymer
M. Dutour Sikirić, A. Sosnik, D. Cohn,H. Füredi-Milhofer
32
22 Čadež Vida
The role of chitin in the biomineralization of the
mollusks and its potential as a bone tissue replacement
biomaterial
Čadež, Vida; Dutour Sikirić, Maja; Šegota, Suzana;
Domazet Jurašin Darija; Pletikapić, Galja; Škapin, D. Srečo;
Sondi, Ivan
33
23 Hélène
PHILIPPOT
Bacteriophage loading of calcium phosphate bio-
ceramics: a tool against infections in bone surgery
Hélène PHILIPPOT, Jean-Christophe HORNEZ, Edwige
MEURICE*, Shaan CHAMARY, Amira HANAFIA*, Anne
LERICHE, Franck BOUCHART
34
24 Maliheh Amini
Moghaddam
Formation of polymer particles with
supercritical CO2: A review
Maliheh Amini Moghaddam
35
25 Stoplova Petra The Optimization of Method for Preparing
Microparticles
Stoplova Petra, Sedlarik Vladimir
36
26 Petr Stloukal The effect of plasma hydrophobization on the realase of
Temozolomide from polyester and polylactic acid
Petr Stloukal, Vladimír Sedlařík, Ivan Chodák, Igor Novák
37
27 Salamanna
Francesca
Strontium substituted and alendronate functionalized
hydroxyapatite nanocrystals in an ovariectomized rat
spinal arthrodesis model
Salamanna F, Giavaresi G, Parrilli A, Torricelli P, Boanini
E, Bigi A, Nicoli Aldini N, Fini M
38
28 Liga Stipniece
Development of nano-hydroxyapatite/poly(vinyl alcohol)
hybrid systems for prospect drug delivery applications
Liga Stipniece, Vitalijs Rjabovs, Inga Narkevica, Kristine
Salma-Ancane, Janis Locs
39
29 Hrabalikova
Martina Nisin immobilization into polyester based microspheres
Hrabalikova Martina, Sedlarik Vladimir 40
11
Serial
Number Authors Name Title
Page
Number
30 Kucharczyk
Pavel
Polyanhydride / wax system for controlle co-release of
bioactive compounds.
Kucharczyk Pavel, Sedlarík Vladimír
41
31 Virginija
Jankauskaitė
The analysis of structure of electrospun nonwoven
material with hydroxyapatite particles
Evaldas Bolskis, Virginija Jankauskaitė, Erika
Adomavičiūtė, Sigitas Stanys, Justė Baranauskaitė, Jurga
Bernatonienė, Onder Albayrak
42
32 Inga Narkevica
Nanostructured Composites Based on ɛ-Polylysine and
Hydroxyapatite for Bone Tissue Repair
Inga Narkevica, Liga Stipniece, Kristine Salma-Ancane,
Janis Locs, Jurijs Ozolins
43
33 Luis Zelaya-
Lainez
The allocation of bone fluid in bovine cortical bone
utilizing a multi-technique analysis
Luis Zelaya-Lainez, Hawraa Kariem, Winfried Nischkauer2,
Andreas Limbeck, Christian Hellmich
44
Invited Speakers
Oral Presentation
Poster Presentation
12
SEARCHING FOR NEW MATERIALS FOR BONE IMPLANTS: CHALLENGES
AND SOLUTIONS OF IN VITRO BIOCOMPATIBILITY ASSESSMENT
Radostina Alexandrova1*, Tanya Zhivkova1, Lora Dyakova2, Boyka-Andonova-Lilova1, Abedulkadir
Abudalleh1, Milena Georgieva3, Georgi Miloshev3, Nabanita Saha4, Diana Rabadjieva5, Stefka
Tepavitcharova5 1Institute of Experimental Morphology, Pathology and Anthropology with Museum, Bulgarian Academy
of Sciences, Sofia, Bulgaria
2Institute of Neurobiology, Bulgarian Academy of Sciences, Sofia, Bulgaria
3Institute of Molecular Biology, Bulgarian Academy of Sciences, Sofia, Bulgaria
4Center of Polymer System, Tomas Bata University, Zlin, Czech Republic 5Institute of General and Inorganic Chemistry, Bulgarian Academy of Sciences,
Sofia, Bulgaria
Abstract
Improving the quality of life is a basic priority in EU policies. Among the diseases with most serious
clinical consequences and economic burden are these associated with bone defects.
Due to the disadvantages and limitations of autografts (e.g. limited availability, additional surgical
trauma) that are still considered as clinical "gold” standard and allografts (limited availability, foreign
body immune reactions, risk of infections) there is a growing need for the development of new artificial
bone substitute materials with improved physicochemical, mechanical and biological properties as close
as possible to those of natural bone, that has to be substituted.
The aim of our study was to evaluate the influence of ion-modified calcium phosphates and brushite
bone cements based on amorphous calcium phosphate and tartaric acid on cell viability and
proliferation.
The following cell cultures were used as model systems: i) human (Lep3, MRC5, BJ) and mouse
(BALB/c 3T3, L929) fibroblasts; ii) mouse bone marrow cells; iii) primary cultures from mouse bone
explants; iv) some human permanent cancer cell lines. The investigations were performed by direct (the
cells were cultured on the material’s surface) and indirect (cells cultured in material’s “extracts”)
experiments using methods such as MTT test (the “Gold” standard for cytotoxicity evaluation), neutral
red uptake cytotoxicity assay, crystal violet staining, trypan blue dye exclusion technique; double
staining with acridine orange and propidium iodide; single cell gel electrophoresis (Comet assay), light
and electron microscopy.
The results obtained reveal that the application of the above mentioned complex approach based on
methods with different molecular/cellular targets and mechanisms of action and various experimental
models (each one with its own advantages and limitations) is suitable and highly informative for in vitro
biocompatibility assessment of new materials for bone implants. Although the preliminary data
obtained for some of the examined materials are promising, additional investigations are required to
clarify better their biocompatibility, osteoconductivity and osteoinductivity
Acknowledgement
This work was supported by Fund “Scientific Research”, Bulgarian Ministry of Education and Science
(Grunt № T 02 5/12.12.2014 and Grant № Б 02 30/12.12.2014) and by COST Action MP 1301.
13
BIOMATERIALS’ ELICITED MESSAGES INFLUENCE EPIGENETICS
Yannis F.Missirlis
University of Patras
Abstract
All living entities, including cells, respond to their environmental cues in ways that we only slightly
know. In tissue engineering applications cells interrogate their environment, consisting of chemical,
topographical and other physical cues, like mechanical signals. Evidence is being built up slowly that
external mechanical signalling to cells are transmitted all the way to the chromatin, where they
deconvolute the highly convoluted structure to facilitate relevant biochemical reactions that result in
changes of the cell’s epigenome. Scant evidence to this phenomenon will be presented.
14
STRUCTURALLY AND FUNCTIONALLY GRADED BIOCOMPOSITES AND
SCAFFOLDS BASED ON CA-PHOSPHATES FOR BONE TISSUE
ENGINEERING
Cihlar J.*, Drdlik D., Novotna L.,Castkova K., Cihlar J. jr,
CEITEC - Central European Institute of Technology, Brno University of Technology, Purkynova
656/123, 612 00, Brno, Czech Republic
Abstract
This contribution presents some results of a study of phase composition, morphology, mechanical and biological
properties of functionally gradient composites and scaffolds in particular based on Ca-phosphate modified by
doped zirconia or silica obtained during solution the project LD14072 (MEYS CZ) involved in the COST Action
MP1301.
A. The influence of the grain size of alumina and zirconia ceramics on the spreading and adhesion of MG63 cell
lines was first investigated. The highest cell spreading and adhesion was obtained for ZrO2 ceramics with an
average grain size of 100 nm. The cell selection was observed on layered ZrO2/Al2O3 composites. The cells
predominantly adhered to ZrO2 layers. The results showed a positive influence of nanostructured ceramic
surfaces on biological behaviour of MG63 cells.
B. Mixtures of nanoparticles of hydroxyapatite nanoparticles ZrO2 doped with CaO and Y2O3 were prepared by
precipitation synthesis and hydrothermal treatment. Analysis of phase composition of the sintered ceramic
composites showed decomposition of hydroxyapatite to tricalcium phosphates and partial transformation of
ZrO2 to CaZrO3. Modulus of elasticity and strength of sintered HA and ZrO2 composites were affected by
porosity. The bioactivity of HA/Ca-ZrO2 composites was higher than HA/Y-ZrO2 ones.
C. Electrophoretic deposition was used for preparation of composite layer containing HA and microfibers of
tetragonal ZrO2. During sintering occurred decay of HA to -TCP, β-TCP and calcium phosphate partially
substituted with Li+ cations. The fracture toughness of the composites was twice as high as toughness of pure
HA due to oriented t-ZrO2 microfibers, finer microstructure, tough β-TCP phase and pores presented in the
composites. Composites showed bioactive behavior.
D. Porous composite scaffolds consisted of bioinert alumina toughned zirconia (ATZ) core and bioactive surface
layer based on Ca-phosphates. The core was prepared by replication technique and the surface layer by dip-
coating. Maximum strength had scaffolds with the core from 95 wt.% ZrO2 and 5 wt.% Al2O3. When tested in
simulated body fluid an apatite layer formed on coated scaffold surfaces and none of composite materials had
cytotoxic behavior when tested with a cell line MG63.
E. The effect of silica on the mechanical and biological properties of the macroporous bioceramic scaffolds based
on calcium phosphate and SiO2 prepared by replication technique was studied. During sintering was observed
a decomposition of hydroxyapatite to alpha and beta tricalcium phosphates partially substituted by silicate
ions. The presence of cristobalite SiO2 phase had positive effect on the compressive strength of scaffolds,
which reached up to 30 MPa. In the course of test of scaffolds in simulated body fluid the apatite layer formed
on the scaffold surfaces containing 0 to 20% cristobalite. In vitro tests using stem cells (ADSC) indicated that
cells were viable and TCP/SiO2 composite was bioactive.
F. When solving the project the methods of shaping of bioceramic layers and scaffolds were inovated: EPD
method, template method and the method of in situ foaming. Currently, was launched research into 3D
printing of bioceramic scaffolds using LCM technology on the device CeraFab 7500 (Lithoz), into which we
invite our colleagues from Action NewGen.
15
(PRE-)CLINICAL BONE ENGINEERING ALSO TAKING CO-MORBIDITIES
INTO ACCOUNT
Martijn van Griensven1
Department of Experimental Trauma Surgery, Clinic for Trauma Surgery, Klinikum rechts der Isar,
Technical University of Munich, Ismaninger Strasse 22, 81675 Munich, Germany
Abstract
For bone engineering, constructs of biomaterials, cells and growth factors are most promising.
Biomaterials can be of natural sources, synthetic sources, and composites. The cells used should be
autologous, have the potential to proliferate and to differentiate. Stem cells of different sources can be
obtained, such as from adipose tissue, bone marrow, umbilical cord and amniotic membrane. The age of
the patient plays an important role for the healing success. Moreover, underlying diseases such as
osteoporosis also influence the outcome. Stem cells from different sources were tested. Importantly,
stem cells from young versus old donors were tested regarding their viability, ability to adhere to
different biomaterials and ability to differentiate. Stem cells from young donors have a higher
proliferation rate, are more viable and show more pronounced osteogenic differentiation as measured by
alkaline phosphatase activity and mineral deposition. qPCR measurements partly confirmed these
findings. Differentiation can be influenced by pulsed electromagnetic fields applied for short time
during several days in an interval manner. Specific pulsed electromagnetic fields were able to enhance
proliferation, alkaline phosphatase activity and mineral deposition of mesenchymal stem cells, thereby
enhancing the chance of success of osteogenic constructs. Another possibility for influencing
osteogenesis may be miRNA. miRNA signatures from osteoporotic patients were determined using
miRNA arrays and validated using specific primers. Osteoporotic patients showed 11 miRNA that were
upregulated in serum and 5 of them also in bone tissue. Modifying miRNA expression with antagomirs
influences the osteogenic potential of stem cells positively. Thus, blocking specific miRNAs may
enhance osteogenic differentiation of stem cells on biomaterials. Underlying facts such as age, gender
and disease specific miRNA signatures may influence the interaction of stem cells with biomaterials and
their ability to differentiate. First clinical cases showed enhanced bone healing. This urges us to
continue our efforts in the field of bone engineering in order to improve the quality of care for patients
suffering from impaired bone healing.
16
STIFFNESS AND STRENGTH PREDICTION FOR A HYDROXYAPATITE-
BASED BIOMATERIAL, CONSIDERING BONE REGENERATION
Stefan Scheiner1*, Vladimir R. Komlev2, Alexey N. Gurin3, Christian Hellmich1 1 Institute for Mechanics of Materials and Structures, Vienna University of Technology, Vienna, Austria
2 A.A. Baikov Institute of Metallurgy and Materials Science & Institute of Laser and Information
Technologies, Russian Academy of Sciences, Moscow, Russia
3 Central Scientific Research Institute of Dentistry and Maxillofacial Surgery, Moscow, Russia
Abstract
Bone replacement materials must fulfill various requirements, such as biocompatibility and appropriate
mechanical properties. In particular, the latter include suitable stiffness and sufficient strength. Finding
such so-called biomaterials with adequate mechanical properties based on experimental trial-and-error
approaches is cumbersome and ineffective, thus mathematical is considered a promising complement.
Here, a hydroxyapatite-based, granular biomaterial, developed as bone replacement material with the
human mandible as targeted application region, is studied. This material exhibits a distinctive
hierarchical organization, with different types of pore spaces of characteristic lengths ranging from
nanometers to millimeters. Additionally, once exposed to the targeted physiological environment (i.e.
the immediate vicinity of mandibular bone tissue), new bone tissue forms arou! nd the bi omaterials
granules, whereas the hydroxyapatite crystals undergo resorption. The presented work aims at
estimation of both stiffness and strength of this material, taking into account the compositional changes
due to bone ingrowth and hydroxyapatite resorption. For this purpose, micromechanical
homogenization techniques are employed, yielding a three-step stiffness homogenization scheme,
giving access to the macroscopic stiffness tensor of the scaffold material. On the other hand, the
micromechanical model is extended by a failure criterion relating to the hydroxyapatite needles, based
on which the loading type-specific macroscopic strength of the scaffold material can be deduced. The
above-sketched micromechanical model gives access to stiffness and strength estimates for the scaffold
material, as functions of various material and design parameters of the material, including the macro-
porosity, the granule size, the density of cracks, as well as the rates of bone formation and scaffold
resorption. The presented modelling approach provides insights beneficial both for the scaffold material
design process and for a clinician actually applying such material. Namely, the aforementioned material
and design parameters could be tuned based on model predictions, to optimize the mechanical
properties of the scaffold material. Complementing this way the experimental trial-and-error strategy
that, in this respect, still is the gold standard, would lead to a significant improvement of the design
process.
17
CONTROL OF GELLING KINETICS IN IONOTROPIC POLYMERS
APPLIED TO 3D CELL FRIENDLY ENCAPSULATION
David C Bassett*, Armend G. Håti, Thor B Melø, Bjørn T. Stokke, Pawel Sikorski
Department of Physics, Norwegian University of Science and Technology, Trondheim, Norway
Abstract
Hydrogels are a hugely important class of biomaterial. Interest from the biomedical materials field
originates from the ability of hydrogels, derived from both natural and synthetic origins, to inherently or
be engineered to mimic physicochemical aspects of the extracellular matrix.
Polymers forming hydrogels must be stabilised in a continuous aqueous phase using physical and/or
chemical crosslinks. This may be achieved via strong covalent crosslinks or physical networks
containing transient, reversible junctions or entanglements based on e.g. ionic and hydrogen bonding or
hydrophobic interactions. Of these types, ionic crosslinking offers a convenient route to gelation,
without recourse to covalently modify the polymer, and many biocompatible polyelectrolyte systems,
such as alginate, pectin, gellan and carrageenan, can be cross-linked under physiological or near
physiological conditions. However, ionic gelling kinetics are difficult to control since they are
governed by the intrinsic interactions between the ionotropic polymer and the gelling ion and ionic
diffusion in water. Attempts to slow down the crosslinking process using retardants, or solid reservoirs
of ionic crosslinking agents have been shown to be effective for limited applications, but problems such
as innapropriate gelation times or non-cytocompatible conditions often arise.[1]
In this presentation, control of ionic crosslinking via a novel approach based on tuning the availability
of the gelling ions will be described. This approach enables the preparation of two stable polymer
solutions which upon mixing react to form a crosslinked hydrogel.[1] The time course over which this
process occurs can be tuned from a few seconds to tens of minutes, thus representing a highly versatile
system. All reactants are contained in the aqueous state and cell toxicity studies indicate that this
approach is highly cytocompatible. Therefore this has particular utility to otherwise challenging
applications in biocompatible 3D soft material structuring using, for example, microfluidics and
additive manufacturing. By way of demonstration of this, high viability cell encapsulation using this
novel method via microfluidic generated droplets will be presented.[2]
References 1. Bassett DC, Håti AG, Melø TB, Stokke BT, Sikorski P. Competitive ligand exchange of
crosslinking ions for ionotropic hydrogel formation. J. Mater. Chem. B. 2016; 4: 6175-6182.
2. Håti AG, Bassett DC, Ribe JM, Sikorski P, Weitz DA, Stokke BT. Versatile, cell and chip friendly
method to gel alginate in microfluidic devices. Lab on a Chip. 2016; 16: 3718-3727.
18
EFFECTS OF X-SHAPED REDUCTION-SENSITIVE AMPHIPHILIC BLOCK
COPOLYMER ON DRUG DELIVERY
Haijun Xiao*, Lu Wang
Address: Centre of Polymer systems, Tomas Bata University in Zlin, nám. T. G. Masaryka 5555
760 01 Zlín, Czech Republic
Abstract
To study the effects of X-shaped amphiphilic block copolymers on delivery and the reduction-sensitive
property on drug release, a novel reduction-sensitive amphiphilic copolymer with a Gemini-like X
shape, (PLGA)2-SS-4-arm-PEG2000 , was successfully synthesised. The formation of micelles was
proved with respect to the blue shift of the emission fluorescence as well as the increase of fluorescent
intensity of coumarin 6 loaded particles. Smaller critical micellar concentration(CMC), smaller size and
higher micellar stability were observed on X-shaped polymer nanomicelles (XNMs). The reduction
sensitivity of polymers was confirmed by the increase of micellar sizes as well as the in vitro drug
release profile. Cytotoxicity assays in vitro revealed that the blank XNMs were nontoxic against A2780
cells up to a concentration of 50 μg/mL, displaying good biocompatibility. DTX loaded X-shaped
polymer nanomicelles (DTX/XNMs) were more toxic against A2780 cells than other formulations in
both dose- and time-dependent manners. Cellular uptake assay displayed a higher intracellular drug
delivery efficiency of XNMs than that of nanomicelles prepared with linear polymers. Besides, the
promotion of tubulin polymerisation induced by DTX was visualised by immunofluorescence analysis.
Therefore, this X-shaped reduction-sensitive block copolymer based on PLGA and PEG could
effectively improve the micellar stability and significantly enhance the therapeutic efficacy of DTX by
increasing the cellular uptake and selectively accelerating the drug release inside cancer cells.
19
“BACTERIAL CELLULOSE” FROM APPLE JUICE MEDIUM: A
SUSTAINABLE RAW MATERIAL FOR HYDROGEL
Smarak Bandyopadhyay, Zandraa Oyunchimeg, Lenka Jlinkova, Nabanita Saha* and Petr Saha
Centre of Polymer Systems, University Institute, Tomas Bata University in Zlin,
třída Tomáše Bati 5678, 760 01 Zlin, Czech Republic
Abstract
In this study, our approach is to establish an inexpensive nutrient medium for optimal production of
bacterial cellulose1. In the Europe, apple production is usually high and was the highest produced fruit
in 2015. But, enormous amount of non-edible apples are wasted every year in Czech Republic and
Europe annually due to sour taste or high acidic content2. Hence, emphasis has been given on “Apple”
to launch it as one of the cheap resource of nutritive medium for the production of Bacterial Cellulose
(BC); a sustainable raw material for Hydrogel which has synergistic effect in Hydrogel formation. BC
production was conducted at 30ºC, incubated for 15 days under static state using 250 ml conical flask
where, 100 ml sterile apple juice (pH 3.8-4.3), was used to supply nutrient for bacterial culture
Gluconobacter xylinum (CCM 3611T) and production of BC. This nature based bio-synthesized
biopolymer (BC) was harvested after 15 days and washed with 0.5N NaOH following the standard
method. A sustainable amount of BC was achieved from the Apple Juice gala medium (AJM). About
4.741gm in weight and 3.3mm in thickness, white; BC membrane was obtained. Moreover, the amount
of production is better in comparison to the HS medium where the mat has an average weight of
1.590gm. It is observed that the cellulose mat has a uniform physical appearance in both “HS medium”
and “AJM”. In conclusion, “AJM” can be considered as a value added and inexpensive substrate for BC
production. The achieved BC can be considered as a cost-effective raw material/bio-material for the
preparation of “Hydrogel”, an elixir medical device / scaffold for bone tissue regeneration.
Acknowledgements
The work was supported by the Ministry of Education, Youth and Sports of the Czech Republic - NPU
Program I (LO1504) and the work done according to the Scientific Program (WG1) of COST
Action_MP 1301_NEWGEN.
Reference
1. 1Rachel T.A. Machado et al. (2016). Komagataeibacter rhaeticus as an alternative bacteria for
cellulose production. Carbohydrate Polymers, 152, 841–849.
2. 2Eurostat news release, 126/ 2016- 22 June 2016.
http://ec.europa.eu/eurostat/documents/2995521/7517627/5-22062016-AP-EN.pdf/8247b23e-f7fd-
4094-81ec-df1b87f2f0bb
20
SYNTHETIC COPOLYMERS FOR BIOMEDICAL NEEDS: FROM
RESORBABLE TO PERMANENT BIOMATERIALS
Karine Salim1*, Eric.Busseron2, Didier Combis1 1 PCAS Head quarter, Longjumeau (Paris region, France).
2 PCAS Expansia, Aramon, France
Abstract
Poly(lactic) acids and poly(lactic-co-glycolic) acids have been of increased interest during the past two
decades as materials for biomedical applications. PLGA are biocompatible and biodegradable, and
exhibit a wide range of degradation timeframes allowing sustained drug release. They also have tunable
mechanical properties for implant applications (tissue engineering and regenerative medicine ) and,
most importantly, are FDA approved polymers.
Moreover, several studiesa has been reported the use of PLGA alone or in combination with other
composite, i.e Hydroxyapatite (HA) material, in nerve & vascular regeneration, cartilage and bone
tissue engineering. The scaffolds combined the degradability of the PLGA with the mechanical support
of HA to form a tissue engineering replacement for bone defects.
In contrast to PLA/PLGA, PEKK (Poly-Ether-Ketone-Ketone) are resistant polymers and non-
biodegradable. PEKK high-performance thermoplastic and biostable copolymers are currently used for
medical applications including permanent surgical implants, dental applications…
Compared to the PEEK (polyether Ether Ketone), the PEKK shows better biocompatibility, wetting,
adhesion and resistance which are major challenges in medical use, laser sintering processes and in
industry (aerospace, automotive, sports, and equipment)b
PCAS proposes a comprehensive spectrum of standards PLA/PLGA, PEKK polymers as well as custom
polymers for the most sophisticated applications
Regulatory aspect (Master File and ISO 10993 guidelines) and quality requirement will also be
discussed.
PCAS, 53 years old is a leading contract manufacturing company in API, Pharmaceutical excipient and
medical device materials. PCAS is a reliable supplier of these biomaterials with over 18 years
experience in producing PLA/PLGA polymers under GMP quality system for parenteral formulations
on the world market.
PCAS can support projects from early phase (R&D) to industrial scale production for marketed product
(multi-kilograms to tons scale), along with worldwide regulatory affairs services.
References aOya KERİMOĞLU et al, ANKEM Derg 2012;26(2) b http://www.arkema.com
For more information: www.pcas.com.
21
CHITOSAN : A VERSATILE TOOL FOR DRUG AND GENE DELIVERY
Antonio Di Martino*
Centre of Polymer Systems, University Institute Tomas Bata University in Zlin, tr.T.Bati
5678,76001,Zlin,Czech Republic
Abstract
Chitosan is a deacetylate derivative of chitin, composed of glucosamine and N-acetyl-glucosamine
units[1]. Its properties, like solubility, biodegradability, biocompatibility, muchoadesivity and
antibacterial are directly affected by the degree of deacetylation (DD) and the molecular weight (Mw)
[2,3].
Since the early 1990s a great interest in chitosan for biomedical application and in particular in drug
delivery has been grown [1]. In contrast to other biodegradable polymers, chitosan exhibits a cationic
character which make it unique. This cationic character is responsible for its success as drug delivery
carrier. Several bioactive compounds which space from anticancer drugs, peptides, antibiotics to
analgesic have been encapsulated, grafted or adsorbed on chitosan particles to control or prolong the
release but also to protect the drug from the degradation, reduce side effects and improve the
therapeutic efficiency in vivo. However, physical and chemical modifcations of chitosan, in case of
neutral environment and loading of hydrophobic drugs, are necessary to improve the performances.
Two chitosan derivatives are mainly obtained; i) hydrophilic, to improve the solubility in neutral and
alkaline media and ii)amphiphilic-hydrophobic, to enhance the encapsulation and release properties.
Next to the delivery of low molecular weight bioactive molecules a great interest to use chitosan as
carrier for polynucleic acids for gene therapy application has been grown and positive results have
been reported in vitro and in vivo [4]. Nucleic acids (DNA and RNA) with any bp number and
conformation can be easily complexed or entrapped in chitosan particles with high loading capacity.
The transfection efficiency depends on formulation factors such as DD and Mw of chitosan, amino-
to-phosphate group (N/P) charge ratio, pH of medium, plasmid, and serum effect [5]. It has been
reported that chitosan-DNA (RNA) polyplexes can be successfully administered by parenteral, oral,
intranasal, pulmonary, and dermal routes.
References
[1] Hu et al., Nanoscale, 5,8, 2013, pp. 3103-3111.
[2] Kofuji et al., European Polymer Journal, 41,11,2005, pp. 2784-2791.
[3] Kean and Thanou, Advanced drug delivery reviews, 62,1,2010, pp. 3-11.
[4] Thibault et al., Molecular biotechnology, 58,10, 2016, pp. 648-656.
[5] Patel et al., World Journal of Medical Genetics, 6,3,2016, pp. 22-33.
22
FUNCTIONAL SIGNIFICANCE OF THREE DIMENSIONAL HYDROGEL
SCAFFOLDS IN BONE TISSUE ENGINEERING – A REVIEW
Probal Basu, Nabanita Saha*, Petr Saha
Centre of Polymer Systems, University Institute, Tomas Bata University in Zlin, Czech Republic
Abstract
Bone is extremely specialized structural framework of human body. One of the exclusive property of
bone tissue is power of regeneration. In past few decades, the bone related disorders has been
increased notably. One of the common bone related disorder is osteoporosis which causes significant
bone fracture. Nearly 75 million people has been affected by osteoporosis in Europe, USA and
Japan[1]. The current treatment protocol involves the application of autograft and allograft or the
administration of inert metallic/ceramic implants. But these methods have been found to insufficient
during severe fracture situation or severe osteoporosis situation. The sole focus of tissue engineering
is the development of three dimensional polymer scaffold which can mimic the extra cellular matrix
(ECM) and facilitate the formation of bone. Hydrogels are three dimensional polymeric cross-linked
network structures which can absorb and retain significant amount of water. The principle function of
the three dimensional hydrogel scaffold material is the appropriate delivery of the seeded cells to the
desired sites, promote cell adhesion, to facilitate the cell differentiation and proliferation. Hydrogel
based biomaterials could efficiently act as a matrices which facilitates tissue regeneration [2].
Composite materials generally mimics the human bone and facilitate the bone development. The
composites have the necessary potential to elevate the efficiency of a bone implant performance.
Research reported that bio-mineralized (CaCO3) PVP-CMC hydrogel could be applied as an
extracellular matrix and might be an efficient approach in bone tissue engineering. This review is an
endeavour to ascertain the functional significance of three dimensional hydrogel scaffold in bone
tissue engineering.
Acknowledgements
This work was conducted within the framework of COST Action MP1301 “New Generation
Biomimetic and Customized Implants for Bone Engineering” (www.cost.eu). The work was supported
by the Ministry of Education, Youth and Sports of the Czech Republic - NPU Program I (LO1504).
References
1. Hernlund E, Svedbom A, Ivergard M, Compston J et. al. (2013) Osteoporosis in the European
Union: Medical Management, Epidemiology and Economic Burden. A report prepared in
collaboration with the International Osteoporosis Foundation (IOF) and the European Federation of
Pharmaceutical Industry Associations (EFPIA). Arch Osteoporos 8:136.
2. Burg KJL, Porter S, Kellam JF (2000). Biomaterial developments for bone tissue engineering.
Biomaterials 21:2347-2359.
23
UTILIZATION OF BIOMIMETIC BIOREACTORS FOR BIOMATERIAL
EVALUATION: GELLAN-GUM HYDROGELS WITH NANO-PARTICULATE
BIOACTIVE-GLASS
Zvicer J.1, Gantar A.2,3, Kocen R.2,3, Veljovic Dj.1, Novak S.2,3, Obradovic B.1* 1Faculty of Technology and Metallurgy, University of Belgrade, Belgrade, Serbia
2Department for Nanostructured Materials, Jožef Stefan Institute, Ljubljana, Slovenia 3Jožef Stefan International Postgraduate School, Ljubljana, Slovenia
Abstract
Biomimetic bioreactors are primarily designed to promote in vitro regeneration of functional tissue
equivalents for tissue engineering (TE) applications by mimicking native cell environments. Yet,
another important role of these bioreactors is to provide reliable characterization of new biomaterials
under physiologically relevant conditions with the ultimate aim to decrease the extent of necessary
animal studies. However, each tissue and organ in the body is exposed to different physical and
biochemical signals, which compels customized approach to the bioreactor design. Hydrodynamic shear
stresses were indicated as a key physical signal enhancing growth of tissue engineered bone in vitro,
while dynamic compressive loadings were shown to affect cell metabolism, structure and biomechanical
properties of articular cartilage both in vitro and in vivo. In the present study, we describe design and
application of two biomimetic bioreactors relevant for bone and cartilage TE. The bioreactor with
medium perfusion is constructed to direct the medium flow directly through the investigated specimen
at physiological velocities while the custom-made bioreactor with dynamic compression provides
dynamic compression of specimens in the physiological regime in conjunction with medium perfusion.
The bioreactors were used to study hydroxyapatite (HAp) formation in gellan gum (GG) scaffolds with
dispersed nano-particulate bioactive-glass (BAG) attractive for use in bone TE. Rehydrated porous
scaffolds (discs 10 mm diameter, 5 mm thick) contained 2 % w/w GG and 2 % w/w BAG (composition:
70 n/n % SiO2, 30 n/n % CaO) while 2 % w/w GG samples served as a control. Samples were
investigated over 14 days in a simulated body fluid (SBF) under (i) continuous perfusion (1.1 ml min-1),
(ii) dynamic compression (5 % deformation, 0.68 Hz frequency, 337.5 m s-1 loading rate, 1 h on/ 1 h
off) and continuous perfusion (1.1 ml min-1), and (iii) static conditions as a control. HAp formation was
examined at the end of experiments by FEG-SEM and RAMAN analyses. Furthermore, mechanical
properties of GG-BAG samples were monitored in the bioreactor with dynamic compression over time.
Results of these studies have shown HAp formation in GG-BAG samples under all conditions, implying
that this process is kinetically controlled. Furthermore, GG-BAG samples preserved structural integrity
over 14 days under physiologically relevant dynamic compression indicating potentials for use as bone
scaffolds.
24
NON-TOXIC BIODEGRADABLE POLYESTER URETHANES BASED ON PLA
Alena Pavelková1*, Pavel Kucharczyk1, Zdenka Kuceková1
Jiří Zedník2 and Vladimír Sedlařík1 1Centre of Polymer Systems, University Institute, Tomas Bata University in Zlin, tr. Tomase Bati
5678, 76001 Zlín, Czech Republic, 2Charles University in Prague, Faculty of Science, Department of Physical and Macromolecular
Chemistry, Hlavova 2030-8, 128 40 Prague 2, Czech Republic.
Abstract
Polylactide (PLA)-based polymers possess indisputable beneficial properties and they are perfectly
convenient for temporary medical applications as drug carriers or resorbable supporting devices.
Nevertheless, PLA is also known for its rigidity and low mechanical properties. On this view, the
fabrication of non-toxic biodegradable material with enhanced mechanical performance is of particular
interest. In the present study, the chain extending reaction using OH- functionalized PLA and
biocompatible lysine diisocyanate can bring an option in the improving of the flexibility of PLA-based
material with a maintenance of biodegradability.
In this research, firstly, the polyethylene glycol (PEG) of various molecular weight is used for the
synthesis of functionalized PLA. Contrary to the different highly demanding synthesis techniques, a
polycondensation reaction is performed, which is economically and environmentally beneficial – no
need for solvents. In the second step, the polymer chains are linked by lysine ethyl ester diisocyanate to
create high molecular weight polyester urethanes. The main objective of this study is to investigate
physico-chemical properties, biodegradability in vitro and cytotoxicity. The results showed that the
molecular weights of the products reached about 50,000 gmol1 and the hydrolytic progress was rapid
in the first 2 weeks; the drop in Mn equaled approximately 70%. Tensile strength testing revealed that
ductility increased alongside reduced molecular weight of PEG. Cytotoxicity tests carried out according
to the EN ISO 10993-5 standard revealed a moderate cytotoxic effect of polymers.
25
MINERALIZATION PROCESS OF CHITOSAN HYDROGELS BY CALCIUM
PHOSPHATE APATITE
S. Ramirez1, 2*, L. David1, T. Delair1, E. Maire2, A. Montembault1, S. Tadier2, L. Gremillard2 1Université de Lyon, Ingénierie des Matériaux Polymères, IMP@Lyon1, CNRS UMR 5223, Université
Claude Bernard Lyon 1, 69622 Villeurbanne Cedex, France 2Université de Lyon, INSA-Lyon, Université Claude Bernard Lyon 1, MATEIS CNRS UMR 5510, 69621
Villeurbanne Cedex, France
Abstract
Bone healing is a field of never ending demand for people of all ages. A bioinspired composite made of
a physical hydrogel of chitosan (CS) mineralized by hydroxyapatite would combine the necessary
healing and angiogenic properties of CS hydrogels [1] with the bioactivity and osteoconductivity of
apatite [2], and present adaptable chemical, architectural, mechanical and biological properties.
Therefore, the objective of this work is to set up a protocol for the mineralization of CS physical
hydrogels and to understand how the precipitation of apatite is influenced by the surrounding organic
phase.
Hydrogels were prepared by dissolution of CS powder in acetic acid and then direct contact with a
NaOH solution, while apatite was precipitated from Ca(NO3).4H2O and (NH4)2HPO4 salts. The
experimental set up developed in this Ph.D. work permitted to investigate two mineralization routes:
- precipitation of an apatite phase inside a preformed hydrogel,
- concomitant apatite precipitation and CS gelation.
X-Ray diffraction, optical and confocal microscopy observations showed the presence of apatite as a
mineral phase on the CS hydrogel surface. Apatite crystals, which were formed, had an imperfect
structure, a low crystalline order similar to the inorganic part of bone tissue.
This work helped to better understand the physico-chemical interactions between CS hydrogel and
calcium phosphate salts. Strategies such as in-situ apatite precipitation and CS gelation are feasible,
opening the way for a gradient of composition and structural properties of the mineral/hydrogel
associations and a platform for additive manufacturing techniques.
Acknowledgement
This work was supported by the LABEX iMUST (ANR-10-LABX-0064) of Université de Lyon, within
the program "Investissements d'Avenir" (ANR-11-IDEX-0007) operated by the French National
Research Agency (ANR).
References
[1] J. Venkatesan and S.-K. Kim, “Chitosan composites for bone tissue engineering-an overview,” Mar.
Drugs, vol. 8, no. 8, pp. 2252–2266, 2010.
[2] K. Lin, C. Wu, and J. Chang, “Advances in synthesis of calcium phosphate crystals with controlled
size and shape,” Acta Biomater., vol. 10, no. 10, pp. 4071–4102, 2014.
26
\
\
\
POLYMER-COATED SILICA PARTICLES FOR CONTROLLED DRUG DELIVERY
Filatova Katerina*, Vladimir Sedlarik
Centre of Polymer Systems, University Institute, Tomas Bata University in Zlin, tr. Tomase Bati 5678,
76001 Zlin, Czech Republic *[email protected]
Abstract
In this work, we have successfully designed and prepared a new stimuli-responsive system that uses
mesoporous silica particles as carriers and polymers as stimuli-responsive coating agents. The combination of
the advantageous structural properties of mesoporous silica particles with provides an efficient strategy for the
preparation of stable and safe drug delivery system. Mesoporous silica particles, with a MCM-41 type
structure, diameter of 187 nm and 4 nm mesopores were synthesized by a modification of Stöber method.
Pristine pH-responsive polymer as well as polymer with polylactic acid were successfully grafted to the surface
of mesoporous silica particles to close the pore entrance avoiding premature release of the loaded cargo and to
provide extended and sustained liberation. The successful grafting of polymer to Si to form hybrid-MSPs was
confirmed by different characterization techniques (TGA, DLS, FTIR, SEM, TEM).
Figure 1. SEM-image of Si carrier and cumulative release of DOX from Si-based carriers, pH=7.
Results showed, that two obvious release stages, a sharp initial burst lasting for 3 hours and a slow release over
a period of 10 days, were observed for the Si without any coating. The hybrid microspheres demonstrated two
distinct drug release stages, a lag stage with no release rate, followed by a sustained release stage, which was
controlled by the degradation of PLA encapsulent. The hybrid structure can deliver a release period of as long
as 5 weeks. More importantly, the initial release burst of the hybrid structure was reduced in comparison with
that of the Si.
Acknowledgements
This work was funded by the Czech Science Foundation (grant No. 15-08287Y) and Ministry of Education,
Youth and Sports of the Czech Republic (grant No. LO1504 and CZ.1.05/2.1.00/19.0409) and Internal Grant
Agency IGA CPS/2016/004.
27
BIOMINERALIZED MAGNETIC HYDROGEL: AN APPLICATION
TOWARDS BIOMEDICINE
Ayan Ray, Nabanita Saha*, Tomas Saha and Petr Saha
Centre of Polymer System, Tomas Bata University, Zlin, Tř. T. Bati 5678, 760 01, Zlin, Czech Republic
Abstract
Magnetic nanoparticles (MNPs) are used in different biomedical and biomedicine applications, whereby
each application requires specific particle properties. To fulfill these requirements, particle properties
have to be optimized by means of variation of crystal structure, particle size, and size distribution 1,2. To
this aim, a novel aqueous coprecipitation method was implemented within the hydrogel matrix for the
formation of MNPs and to achieve biomineralised magnetic hydrogel. It is an emerging and promising
concept of utilizing hydrogel because of its 3 Dimentional, croslinked internal structural properties. In
this work, “PVP-CMC Hydrogel”3 is used as matrix for synthesis for the biomineralized MNP, where
the unique porous networks structure exhibited at freeze dried state. This porous structure embedded
within the hydrogel which function as a chemical reactor, where inorganic salts (FeCl2 and FeCl3) are
reacting with precipitating agents ammonium hydro-oxide (NH4OH) to generate MNP (Fe3O4).
Moreover, as the said hydrogel is prepared with biopolymer, biodegradable polymers and/or co-polymer
like: Polyvinylpyrrolidone (PVP), carboxymethyl cellulose (CMC), polyethelene glycol (PEG), agar
etc. that can be considered as an alternative eco-friendly biomaterial and can be used for versatile
medical application such as a medical device for biomedicine, drug delivery etc. This hydrogel based in
vitro synthesis process can be used to synthesize “bare” MNP, magnetic hydrogel4 and extended to
other oxides by changing the salts for the biomedical applications. Finally, this biomineralized (Fe3O4)
magnetic hydrogel can be recommended for its use as shoe sole for the treatment of foot aimed at
diabetic patient.
Acknowledgement
The work was supported by MŠMT ČR-USA Kontakt II (LH14050) and NPUI (LO1504) Ministry of
Education, Youth and Sports of the Czech Republic. This work was conducted within the framework of
COST Action MP1301 “New Generation Biomimetic and Customized Implants for Bone Engineering”
(www.cost.eu).
References 1. Reddy, L. H., Arias, J. L., Nicolas, J. & Couvreur, P. Magnetic nanoparticles: design and characterization, toxicity
and biocompatibility, pharmaceutical and biomedical applications. Chem. Rev. 112, 5818–5878 (2012).
2. Tran, N. & Webster, T. J. Magnetic nanoparticles: biomedical applications and challenges. J. Mater. Chem. 20,
8760–8767 (2010).
3. Roy, N., Saha, N., Kitano, T. & Saha, P. Novel hydrogels of PVP--CMC and their swelling effect on viscoelastic
properties. J. Appl. Polym. Sci. 117, 1703–1710 (2010).
4. RAY Ayan, SAHA Nabanita, S. T. and S. P. IN SITU SYNTHESIS OF BIOMINERALIZED MAGNETIC
NANOPARTICLE WITHIN HYDROGEL. in 1–6 (Nanocon 2016, 2016).
28
HYALUONAN HYDROGEL/CALCIUM PHOSPHATES COMPOSITES FOR
MEDICAL APPLICATION
Marina Sokolova*, Janis Locs, Dagnija Loca, Ilze Balode
Rudolfs Cimdins Riga Biomaterial Innovations and Development Centre of RTU, Institute of General
Chemical Engineering, Faculty of Material Science and Applied Chemistry, Riga Technical University,
Pulka 3, Riga, LV-1007, Latvia
Abstract
In last decade, fabrication and investigation of inorganic-organic composites based on calcium
phosphates (CaP) and biocompatible polymers has attracted research interest in the field of bone tissue
engineering. Mechanical properties, degradation behaviour, and cell-material interaction are among the
most dominantly affected characteristics resulting from the addition of inorganic material to hydrogel
networks. Hyaluronic acid (HA) is a naturally derived polymer as a major component of the
extracellular matrix, mediating various cellular activities, and has been widely used as tissue
engineering scaffolds. The aim of the current research was to obtain HA/CaP hydrogels using three-
dimensional network formation in inorganic particle suspension and to investigate the influence of
inorganic part on the hydrogel formation and swelling properties.
A homogenous suspension of CaP particles for the preparation of HA and CaP hydrogels (HA/CaP) was
obtained via wet chemical precipitation synthesis.
Hydrogels with HA concentration from 7 – 10 wt% were made under alkaline conditions (0.25 M
NaOH), with 1.4-butanediol diglycidyl ether (BDDE) as crosslinking agent. CaP suspension was added
at HA/CaP (w/w) ratios 2.33, 1 and 0.43, and mixed for 2 hours. Sol-gel transition underwent at 45 °C
for 3 h, followed by sample neutralization with 0.25 M HCl. Swelling behavior of prepared hydrogels
was evaluated in phosphate buffered saline (PBS) at 37 °C.
The effect of HA concentration and HA/CaP ratio on the swelling behavior of the prepared hydrogels
were evaluated. Swelling kinetic studies revealed that the hydrogels swelled rapidly reaching
equilibrium state within 48 hours. Swelling ratio values at equilibrium state in PBS are shown in Fig. 1.
Increasing polymer concentration from 7wt% to 10wt%, the swelling ratios decreased twofold, except
for the sample 0.43 HA/CaP, where hydrogel failed to form with 10wt% HA.
BDDE cross-linked HA/CaP hybrid hydrogels were successfully fabricated with HA/CaP ratio from
2.33 – 0.43. Samples were immersed in PBS solution and it was observed that HA hydrogels maintained
their stable structure for 14 days and HA/CaP hydrogels for 36 days. Furthermore, the amount of CaP
influenced the swelling behavior of obtained hydrogels. Addition of CaP particles to HA hydrogel
reduced swelling ratio for two times.
29
THE EFFECT OF ANTIMICROBIAL AGENTS ON THE STABILITY OF
HYDROGELS
Lucie Nogolova*, Vladimir Sedlarik
Centre of Polymer System, Tomas Bata University, Zlin, Tř. T. Bati 5678, 760 01, Zlin, Czech Republic
Abstract
This work is focused on the effect of antimicrobial agents on the stability of hydrogels. The aim of the
study was the production of new hydrophilic gelling polymer compositions based on acrylic acid
copolymer for application in cosmetology and pharmacy, respectively. The effect of the hydrogel
modification with bioactive compounds, such as nisin (originating from whey fermentation products)
and essential oils of citrus fruit, on physico-chemical and antibacterial properties of the resulting system
was investigated. Prepared hydrogels were subjected to accelerated stress test, during which the pH, the
viscosity and the concentrate of nisin (by RP-HPLC) was measured. The antimicrobial efficacy test was
also carried out. Results showed the addition of nisin decrease pH and viscosity and vice versa the
essential oils do not influence these properties. During the stress test the concentration of nisin was
decreased similarly in all prepared samples. All tested systems proved significant reduction of
microorganisms growth but synergy of nisin and essential oils was found out and it was found to be
promising preservative system.
Acknowledgement
This work was funded by Ministry of Agriculture of the Czech Republic (Project no. QJ1310254) and
the Ministry of Education, Youth and Sports of the Czech Republic (Project no. LO1504).
30
THE DEGRADATION TESTS OF CERAMIC-POLYMER COMPOSITES WITH
HYDROXYAPATITE FOR ORTHOPEDIC IMPLANTS
Sylwia Dąbrowska1,2*, Elżbieta Pietrzykowska1,2, Roman Mukhovskyi1, Agnieszka Chodara1,2,
Tadeusz Chudoba1, Janis Locs4, Witold Łojkowski1,3
1Institute of High Pressure Physics, Sokolowska 29/37, 01-142 Warsaw, Poland 2Warsaw University of Technology, Faculty of Materials Science and Engineering, Warsaw, Poland.
3Bialystok University of Technology, Faculty of Management, Bialystok, Poland. 4Riga Technical University, Riga, Latvia.
Abstract
Bioactive materials which can support bone ingrowth and osseointegration are common used in
orthopedic applications. Bioactive hydroxyapatite (HAP) is one of the inorganic component of hard
tissues, which is manufactured in The Institute of High Pressure Physics of the Polish Academy of
Sciences (IHPP) and it is called GoHAPTM. The morphology, grain size and specific surface area of the
nanopowder can be controlled by the microwave reactor and the high pressure consolidation technology
for ceramic materials [1].
The aim of the GoIMPLANT project was to develop resorbable, tough, strong and biocompatible hybrid
composite implants in according to patient’s needs. To get better mechanical properties in our
laboratory we used combination of GoHAPTM and biocompatibile polymer like polylactic acid (PLA).
We used isostatic pressing and uniaxial pressing to form our hybrid composite in form of cylinder or
wedge. Biodegradable composites can be decomposed naturally after a certain period of implantation
with degraded products, which will stay inside the body. Mechanical and biological performance of
composites for implantation depends on the degradation rate. Used degradation medium, pH and
temperature like inside human body can determine the test. The degradation study of composites was
performed according to the ISO standards of medical devices, using PBS and TRIS-HCL solutions.
Testing the changes of Ca2+ concentration, the conductivity and pH under equilibrium conditions at
37°C PBS was checked once in a week.
Research was realized by the “GoIMPLANT” Project and it is founded by M-Era.Net program of The
National Centre for Research and Development, co-financed from the European Union, Regional
Development Fund.
References
1. Patent application P-369906, Lojkowski et al, The method of nanoplates and method of nanopowder
with nanoplates obtaining from synthetic hydroxyapatite.
31
MECHANICAL AND BIOLOGICAL EVALUATION OF CALCIUM
PHOSPHATE / SILICA BASED MACROPOROUS SCAFFOLDS
Novotna L.1*, Chlup Z.2, Cihlar J. 1 1CEITEC - Central European Institute of Technology, Brno University of Technology, Purkynova
656/123, 612 00, Brno, Czech Republic 2CEITEC - Central European Institute of Technology, Institute of Physics of Materials, Academy of
Sciences of the Czech Republic, Zizkova 22, 616 62, Brno, Czech Republic
Abstract
The aim of this work was to prepare sufficient strong and bioactive scaffolds based on calcium
phosphates and silica. It is generally believed that incorporation of small amount of silica into the lattice
of calcium phosphates has positive effect on both the proliferation of human osteoblasts and the rate of
bone apposition. The effect of crystallized silica on mechanical and biological properties of scaffolds
was therefore studied. Calcium phosphate / silica macroporous scaffolds were prepared by the polymer
replica technique. Scaffolds with different amount of silica (up to 20 wt%) exhibited well-
interconnected open pore structure with spherical pores exceeding 250 µm, the size that allow cells to
penetrate easily through the scaffold. The compressive strength of scaffolds containing silica was higher
than the strength of pure calcium phosphate scaffolds of the same porosity; the strength was similar to
the cancellous bone. The biological evaluations of scaffolds were done by in vitro testing methods. The
apatite formation on the surface of coated scaffolds after immersion in MEM solution indicated a
bioactive behaviour of prepared samples. Neither of tested ceramic material was cytotoxic, thus
scaffolds containing silica could be suitable candidates for clinical applications.
32
DEVELOPMENT OF INJECTABLE BONE SUSBSTITUTE MATERIALS
BASED ON CALCIUM PHOSPHATES AND REVERSE THERMORESPONSIVE
POLYMER
M. Dutour Sikirić1*, A. Sosnik2, D. Cohn2,H. Füredi-Milhofer2
Ruđer Bošković Institute, Bijenička cesta 54, 10000 Zagreb, Croatia1
Casali Institute for Applied Chemistry, The Hebrew University of Jerusalem, Givat Ram Campus,
Jerusalem, Israel2
Abstract
Fully injectable polymers, suitable to be implanted into the human body without surgical procedure may
serve as scaffolds for guided bone regeneration. Among different types of polymers, reverse
thremoresponsive polymer attract special attention due to their ability to form low viscous solutions at
lower temperature, while at higher, i.e. body, temoperature they form gel. Bone induction properties of
such materials could be greatly improved by incorporation of appropriate calcium phosphate crystals.
So far this was attempted by mixing pre-prepared calcium phosphate particles with injectable polymers.
The aim of this investigation was to prepare in situ generated organic-inorganic composites consisting
of reverse thermoresponsive pluronic F127-DMA and in situ precipitated calcium phosphates.
In situ calcium phosphate precipitation was initiated by fast mixing of calcium and phosphate polymer
containing solutions, in water and phosphate buffer. Gel was formed within 5 minutes by raising
temperature to 37oC. Calcium phosphates grown in gel weree characterized by environmental (E-SEM)
and high resolution scanning electron microscopy (HR SEM), electron dispersive spectrophotometry
(EDS) and X-ray powder diffraction (XRD).
Results have shown that finely dispersed slightly aggregated crystalline calcium phosphate particles
were formed, their composition depending on the medium used. In water, calcium
dihydrogenphosphatedihydrate was formed, while in phosphate buffer poorly crystalline apatitic phase
was formed. This points to the simple mode of controlling calcium phosphate phase formed within the
gel.
33
THE ROLE OF CHITIN IN THE BIOMINERALIZATION OF THE MOLLUSKS
AND ITS POTENTIAL AS A BONE TISSUE REPLACEMENT BIOMATERIAL
Čadež, Vida1*; Dutour Sikirić, Maja1; Šegota, Suzana1; Domazet Jurašin Darija1; Pletikapić, Galja1;
Škapin, D. Srečo2; Sondi, Ivan3
Ruđer Bošković Institute, Bijenička cesta 54, 10000 Zagreb, Croatia1
Advanced Materials Department, Jožef Stefan Institute, Jamova cesta 39, 1000 Ljubljana, Slovenia2
Faculty of Mining, Geology and Petroleum Engineering, Pierottijeva 6, 10000 Zagreb, Croatia3
Abstract
Functionalized hydrogels have a great potential as a biomaterial for bone tissue replacement. They can
be used as matrices for bone tissue ingrowth, as a therapeutics (by increasing disk hydrations) or/and as
a drug delivery systems. The challenge is to develop hydrogel scaffolds that are highly porous, but
strong enough to withstand diverse mechanical strains, while mimicking the natural extracellular matrix
(NEM) structure. Integration of natural extracellular organic matrix as a part of a biomaterial provides
simple and efficient way to obtain scaffolds with enhanced framework structure. In this sense, marine
organisms attract special attention as NEM sources. Their extracellular organic matrix is rich with
diverse proteins and chitinous fibers. Potential of chitin is already known in the biomimetics1, but most
research is conducted on the chitin isolated from arthropods, i.e. on -chitin. In comparison, -chitin is
more suitable to serve as a biomaterial since it has better water retention possibilities, but it is also more
difficult to extract.2 For example, it can be found in most biominerals of mollusks, but they are more
mineralized than arthropods, so its extraction can be demanding and detrimental to chitin.3,4
The aim of this research was to determine which mollusk, the cuttlefish (Sepia officinalis, L.) or the
shellfish (Arca noae, L.), is more suitable source of chitin. Structure – function relationships of organic
matrix and biomineral structures were examined from both, morphological and structural perspective
with the use of Fourier transmission spectroscopy (FTIR), atomic force microscopy (AFM) and field
emission scanning electron microscopy (FESEM). Obtained results indicate that the process of isolation
is much more feasible in the cuttlefish and that isolated chitin has a good structure and properties to be
used as scaffold for functionalized hydrogels.
References
1.Jayakumar, R. et al. Chitin Scaffolds in Tissue Engineering. Int. J. Mol. Sci. 12, 1876–1887 (2011).
2.Younes, I. & Rinaudo, M. Chitin and Chitosan Preparation from Marine Sources. Mar. Drugs 13,
1133–1174 (2015).
3.Gotliv, B.-A., Addadi, L. & Weiner, S. Mollusk Shell Acidic Proteins: In Search of Individual
Functions. ChemBioChem 4, 522–529 (2003).
4.Rocha, J. H. G. et al. Scaffolds for bone restoration from cuttlefish. Bone 37, 850–857 (2005).
34
BACTERIOPHAGE LOADING OF CALCIUM PHOSPHATE BIO-
CERAMICS: A TOOL AGAINST INFECTIONS IN BONE SURGERY
Hélène PHILIPPOT, Jean-Christophe HORNEZ, Edwige MEURICE*, Shaan CHAMARY, Amira
HANAFIA*, Anne LERICHE, Franck BOUCHART *
LMCPA, Pôle Universitaire de Maubeuge, boulevard Charles de Gaulle, 59600, MAUBEUGE
* LMCPA, Pôle Universitaire de Cambrai, 6 rue de Rambouillet, 59400, CAMBRAI
Abstract
Calcium phosphate ceramics, particularly hydroxyapatite and tri-calcium phosphate, have been used
for several decades as choice materials for bone filling in bone and joint surgery, because of their
mineral composition, and very good bio-activity.
Bacterial infections are a major complication of these surgical procedures, and their treatment with
antibiotics is frequently inefficient because of their poor diffusion in the bone.
These nosocomial infections have become more and more frequent in recent years, because the
number of patients requiring such procedures is on a constant rise (as the population gets older), but
also because of the increase in the apparition of antibiotic-resistant bacterial strains.
The aim of this project is to address both of these issues, by using ceramics with porous structures,
combined or not with organic polymers, in order to load them with antibacterial agents and to ensure
their controlled release over time. Moreover, as the use of antibiotics has shown some limitations,
using some other form of antibacterial therapy is becoming essential. Phage therapy has the
advantages of having a fast and discriminating action on specific bacterial strains, which reduces the
occurrence of side effects, and dramatically slows down the apparition of new resistant strains. It can
be used with relative ease as a prophylactic.
In this context, the LMCPA as demonstrated the natural capacity of calcium phosphate ceramics to
retain and release bacteriophages for up to 6 days. Their ability to retain the phages, and their release
behavior varies with the amount of micropores in the ceramic. The overall goal of this project is to
develop new devices that can administer phage based drugs locally, to prevent and cure infections
related to bone and joint surgery.
35
FORMATION OF POLYMER PARTICLES WITH
SUPERCRITICAL CO2: A REVIEW
Maliheh Amini Moghaddam1
Tomas Bata University in Zlín, Centre of Polymer Systems, Av. Tomas Bata 5678
760 01 Zlín
Abstract
Recently some particle formation processes and micronization has been increased due to growing
concerns regarding health, safety and the environment. The particle formation processes are either
solvent-less or use environmentally acceptable solvents such as carbon dioxide have come into favor.
Supercritical CO2 (sc CO2) (T > 31.1 °C, P > 7.3 MPa) has been used in pharmaceutical industries to
minimize the use of organic solvents and micronize (0.1–5 mm) pharmaceuticals. Control of particle
size increases the dissolution rate of drugs into the body. The low viscosity and high diffusivity of
scCO2 offer the possibility of novel processing routes for drug delivery. There are several techniques
for the particle formation which is used supercritical CO2 either as a solvent, and solution is rapidly
expanded (rapid expansion of supercritical solutions, RESS), or as a solute leading to the formation of
particles from gas-saturated solutions (PGSS), or as an anti-solvent leading precipitation induced
particle formation as in (supercritical anti-solvent (SAS),gas anti-solvent (GAS) solution enhanced
dispersion by supercritical fluid (SEDS) and aerosol solvent extraction systems (ASES)).
36
THE OPTIMIZATION OF METHOD FOR PREPARING MICROPARTICLES
Stoplova Petra*, Sedlarik Vladimir
Centre of Polymer Systems, University Institute, Tomas Bata University in Zlin, tr. Tomase Bati 5678,
76001 Zlin, Czech Republic
Abstract
Alginate is a natural polysaccharide which is obtained from brown algae. Alginates are widely used in
food industries. It is also used in pharmaceutical industries as a matrix for the entrapment or
encapsulated drugs. Alginate particles can be produced in several ways. One of them is a spray-
coagulation method. The particles produced by this method are porous. Calcium chloride solution is
used as gelation agent in various concentrations. This study is focused on optimalization of method for
preparing alginate particles mainly on their size and shape. Conditions that are changed are flow rate, air
pressure, height of needle, concentration of alginate and CaCl2 solution. The shape and a surface of
particles is identified from scanning electron microscopy (SEM). Their size is meassured by
Mastersizer 3000 laser particle size analyzer. Results showed that changed conditions have an effect on
physical properties of particles. The size of particles is decreased with decreasing flow rate, increasing
pressure of air and also with more concentrated solution of calcium chloride.
Acknowledgement
This work was funded by the Ministry of Argiculture of the Czech Republic (Project no. QJ1310254)
and the Ministry of Education, Youth and Sprots of the Czech Republic (Project no. LO1504).
37
THE EFFECT OF PLASMA HYDROPHOBIZATION ON THE REALASE OF
TEMOZOLOMIDE FROM POLYESTER AND POLYLACTIC ACID
Petr Stloukal1, Vladimír Sedlařík1, Ivan Chodák2, Igor Novák2
1 Centre of Polymer Systems, Tomas Bata University in Zlín, nam. T. G. Masaryka 5555, 760 01 Zlín,
Czech Republic.
2 Polymer Institute, Slovak Academy of Sciences, Dúbravska cesta 9, 845 41, Bratislava 45, Slovakia
Abstract
In the study, the effect of plasma deposited siloxane coating on the release kinetics of drug
Temozolomide (TMZ) from the thin biodegradable polyesters films: polylactic acid (PLA) and
polyester urethane (PEU) were investigated. The changes in surface properties were studied by the
method of water contact angle measurements, nanoindentation and X-ray photoelectron spectroscopy.
The hydrolytic degradation of materials as well as release kinetics of TMZ was investigated by using
chromatographic and spectroscopic techniques, respectively. The plasma deposited hydrophobic
siloxane coating significantly modified the release kinetics of TMZ from both polymers especially from
PLA. The initial burst effect and the release rate of TMZ was significantly reduced. The hydrolytic
degradability of materials were slightly reduced after hydrophobization especially for PLA as an effect
of wettability modification.
Acknowledgements
This work was financially supported by the Czech Science Foundation
(Grant No. 15-08287Y) and the Ministry of Education, Youth and Sports of the Czech Republic (grant
No. LO1504). The work was also supported by the Ministry of Education, Science, Research and Sport
of the Slovak Republic and Slovak Academy of Sciences, project VEGA 02/0199/14.
38
STRONTIUM SUBSTITUTED AND ALENDRONATE FUNCTIONALIZED
HYDROXYAPATITE NANOCRYSTALS IN AN OVARIECTOMIZED RAT SPINAL
ARTHRODESIS MODEL
Salamanna F1-*, Giavaresi G1,2, Parrilli A1, Torricelli P1,2, Boanini E3, Bigi A3, Nicoli Aldini N 1,2, Fini M1,2.
1 Laboratory of Biocompatibility, Technological Innovations and Advanced Therapies-Department RIT
Rizzoli-Rizzoli Orthopedic Institute, Bologna, Italy;
2 Laboratory of Preclinical and Surgical Studies, Rizzoli Orthopedic Institute, Bologna, Italy;
3 Department of Chemistry “G. Ciamician”, University of Bologna, Italy *[email protected]
Abstract
Advanced age, osteoporosis and unhealthy life-style diminish bone formation and remodeling rate so fusion
enhancement techniques could be beneficial for patients with osteoporosis who need spinal arthrodesis.
Herein we investigated the posterolateral fusion rate in ovariectomized (OVX) rats using two new materials:
strontium substituted hydroxyapatite (SrHA) nanocrystals and alendronate functionalized HA (HA-AL)
nanocrystals. HA was synthesized at increasing Sr concentrations (SrHA5; SrHA10) and alendronate
content (HA-AL7; HA-AL28). A posterolateral spinal fusion model in twenty-five Sham-Operated and in
twenty-five OVX female rats was used. Materials were bilaterally implanted between transverse processes of
lumbar vertebrae. Sham and OVX animals were divided in five groups: HA, SrHA5, SrHA10, HA-AL7 and
HA-AL28. The assessment of bone fusion was carried out by micro-CT, histology and histomorphometry.
Some gaps between the transverse processes were observed by micro-CT in OVX HA group, while they
were not present in the other groups. These results were consistent with histological and
histomorphometrical analyses showing that in OVX animals SrHA and HA-AL materials displayed
significantly higher BV/TV and Tb.Th and significantly lower Tb.N and Tb.Sp in comparison with HA
alone. Results suggest that in spinal fusion the incorporation of Sr, as well of AL, improves the biological
performance of HA in a dose dependent way, and represents a promising strategy especially in osteoporosis
patients with high risks of spinal fusion failure. Results also suggest that HA-AL28 could be the candidate
biomaterial for spinal fusion in osteoporotic subjects.
39
DEVELOPMENT OF NANO-HYDROXYAPATITE/POLY(VINYL ALCOHOL)
HYBRID SYSTEMS FOR PROSPECT DRUG DELIVERY APPLICATIONS
Liga Stipniece1*, Vitalijs Rjabovs2, Inga Narkevica1, Kristine Salma-Ancane1, Janis Locs1 1Rudolfs Cimdins Riga Biomaterials Innovations and Development Centre of RTU, Institute of General
Chemical Engineering, Faculty of Materials Science and Applied Chemistry, Riga Technical University,
Pulka 3, Riga, LV 1007, Latvia 2Institute of Technology of Organic Chemistry, Faculty of Materials Science and Applied Chemistry,
Riga Technical University, Paula Valdena 3/7, Riga, LV-1048, Latvia
Abstract
Fabrication and investigation of inorganic-organic composites based on calcium phosphates (CaP) and
biocompatible polymers has attracted research interest in the field of bone tissue engineering for
potential application in developing of controlled release drug delivery systems. Hydroxyapatite (HAp)
is used for bone-tissue engineering, due to its bioactivity and osteoconductivity. Poly(vinyl alcohol)
(PVA) is a hydrophilic polymer possessing biocompatibility. In order to achieve the synergic effect of
bioactivity of HAp and adjustable biodegradability of PVA, PVA/HAp microgranules were formed by
spray drying process of the polymeric suspensions containing HAp particles synthesized in situ.
Moreover, PVA was modified with succinic anhydride to introduce carboxylic acid residues that either
can be modified with organic biologically active compounds or can chelate bioactive metallic ions on
the surface of the HAp particles.
PVA was modified to introduce –COOH groups by reaction between the –OH groups and succinic
anhydride. The composites were obtained using two different preparation techniques: (a) aqueous
solutions of the pure or modified PVA (M-PVA) and as-synthesized HAp suspension were mixed; (b)
HAp was synthesized in situ, i.e., in the presence of aqueous solutions of the PVA or M-PVA. In both
cases, HAp were synthesized from Ca(OH)2 and H3PO4 through aqueous precipitation. In order to yield
moldable composite powders, the suspensions were spray dried at temperature under boiling point of
PVA. The characteristics of the products such as morphology, chemical and phase compositions etc.
were studied.
The feasibility of producing microgranules (~2-10 μm) composed of nanosized HAp crystallites
homogenously embedded in the polymer matrix opens the possibility of developing injectable bone
fillers or formulation of bone cements with hydraulic setting. The innovative biomaterial could be
employed for controlled release of therapeutic agents at wound sites. This preliminary research
contributes to further biomedical applications.
Acknowledgements
This work has been supported by the National Research Programme No. 2014.10-4/VPP-
3/21“MultIfunctional Materials and composItes, photonicS and nanotechnology (IMIS2)” Project No. 4
“Nanomaterials and nanotechnologies for medical applications”.
40
NISIN IMMOBILIZATION INTO POLYESTER BASED MICROSPHERES
Hrabalikova Martina*, Sedlarik Vladimir
Centre of Polymer Systems, University Institute, Tomas Bata University in Zlin, Tr. Tomase Bati 5678,
76001 Zlin, Czech Republic
Abstract
The present study deals with the development of the polyvinyl alcohol (PVA) cross-linked with glutaric
acid (GA) core and polylactic acid (PLA) multi-hollow shell encapsulation systems that could provide
the effective encapsulation of hydrophilic active agents in the hydrophobic polymer matrix with
controlled morphology. The microsphere systems were used as delivery systems for immobilization of
model antibacterial agent – peptide nisin (NIS). The effect of cross-linking and the NIS amount on their
morphology was investigated by using scanning electron microscopy (Figure 1). Encapsulation
efficiency and release profile of nisin from the microspheres were studied by high performance liquid
chromatography. Antibacterial activity of the prepared systems was tested by dilution and spread plate
technique. Results showed the microspheres in the size range of 9-16 µm in diameter with the spherical
multi-hollow core-shell structure. The presence of cross-linking agent GA influences the release profile
of the NIS and has synergistic effect on Listeria monocytogenes grown reduction.
Figure 1 SEM pictures of PLA/PVA (A), PLA/PVA/NIS (B), PLA/PVA/GA/NIS (C)
Acknowledgements
This work was funded by the Ministry of Agriculture of the Czech Republic (Project no.
QJ1310254) and the Ministry of Education, Youth and Sports of the Czech Republic (Project no.
LO1504).
41
POLYANHYDRIDE / WAX SYSTEM FOR CONTROLLE CO-RELEASE OF
BIOACTIVE COMPOUNDS
Kucharczyk Pavel1*, Sedlarík Vladimír2 1Centre of Polymer Systems, University Institute, Tomas Bata University in Zlin, tr. Tomase Bati 5678,
76001 Zlín, Czech Republic,
Abstract
Polyanhydrides are less known types of biodegradable polymers in contrast with e.g. polylactides. Due
to the presence of unstable anhydride bonds these polymers are highly suspeccible to hydrolysis and
consequently fast degradation. In combination with, in most cases, very hydrophobic nature, majority of
polyanhydrides exhibit so called “surface erosion” degradation pattern. This is highly desirable in drug
release applications because zero order release kinetic can be achieved. On the other hand, their fast
degradation can be problematic in cases in which long term application is required.
This work describes application of biodegradable poly(sebacic anhydride) (PSA) based matrix
combined with biocompatible bees wax for co-release of two model drugs acetaminophen (AAP) and
ibuprofen (IBU). The studied blend system consist between 0-75% of wax and fixed drug in
concentration 5 mg.g-1. The wax in these systems should work as water repelling agent to slow
degradation rate and prolong drug release. The blends were prepared by mixing all components in
chloroform followed solvent evaporation and preparation of tablets (13mm diameter, 2mm thickness)
by cold press moulding. The release experiment was performed into phosphate buffer (pH 7.4) at 37°C
and amount of released drug was detected by HPLC with UV detection.
It was found that presence of wax in PSA matrix can significantly slow down the release rate of both
model drugs as well as the initial “burst” effect phenomenon.
42
THE ANALYSIS OF STRUCTURE OF ELECTROSPUN NONWOVEN
MATERIAL WITH HYDROXYAPATITE PARTICLES
Evaldas Bolskis1*, Virginija Jankauskaitė1, Erika Adomavičiūtė1, Sigitas Stanys1, Justė Baranauskaitė2,
Jurga Bernatonienė2, Onder Albayrak3 1 Kaunas University of Technology, Kaunas, Lithuania
2Lithuanian University of Health Sciences, Kaunas, Lithuania 3 Mersin University, Mersin, Turkey.
E-mail: [email protected]
Abstract
Synthetic biodegradable polymers attracted special attention in electrospinning due to the elimination of
a second surgery to remove the implanted carrier. Electrospinning is a process by which ultrafine fibers
with nanometer or submicrometer diameters from different polymers (polyvinyl alcohol,
polycaprolactone, polylactide acid, chitosan, etc.) can be produced. Electrospun fibers exhibit several
unique characteristics, such as large surface area to mass or volume ratio, small pore size between
depositing fibers of the electrospun mat.
Hydroxyapatite (Ca10(PO4)6(OH)2) (HA) is a biomimetic material and are widely used in biomedical
applications in combination with different materials since it is an inorganic component of bones and
teeth. Due to its poor mechanical properties it cannot be used alone as bone implant material.
The aim of this study was to estimate the influence of HA particles surface treatment on the diameter of
electrospun polyvinyl alcohol fibers. Additionally, the possibility to use methacrylic acid/methyl
methacrylate copolymer for electrospinning process and fibres structure was investigated.
Two kinds of electrospun mats from polyvinylachol (PVA) and biodegradable methacrylic acid–methyl
methacrylate (MA/MMA) copolymers were used for investigations. Electrospinning solution of 12%
concentration were prepared by dissolving of PVA (Mw = 72000 g/mol) in distilled water at 80 °C
temperature. Electrospinning solution of 5–10% concentration was prepared by dissolving MA/MMA
(Eudragit E100) in ethanol. Undoped HA or boron doped HA (BHA) powders were added (5 wt% by
weight) to PVA solution.
It was obtained that HA particles treatment procedure influenced on the diameter of electrospun PVA
nano-microfibers. The doping of HA particles by boron decrease amount of nanofibers with diameter of
less than 100 nm.
Preliminary investigations show that biodegradable MA/MMA copolymer can be used as support
(delivery) material for HA particles.
43
NANOSTRUCTURED COMPOSITES BASED ON Ɛ-POLYLYSINE AND
HYDROXYAPATITE FOR BONE TISSUE REPAIR
Inga Narkevica*, Liga Stipniece, Kristine Salma-Ancane, Janis Locs, Jurijs Ozolins
Rudolfs Cimdins Riga Biomaterials Innovations and Development Centre of RTU, Institute of General
Chemical Engineering, Faculty of Materials Science and Applied Chemistry, Riga Technical University,
Riga, LV-1007, Latvia
Abstract
The development of biomaterials for tissue engineering is to create surfaces which can provoke specific
cellular responses and direct new tissue regeneration. Thus, inorganic/organic systems such as calcium
phosphate/natural biopolymers, have attracted a great deal of attention over the past few years as a
carrier for the delivery of antibiotics in the treatment of bone tissues. Nanocrystalline hydroxyapatite
(HAp) is a major inorganic component of bone with excellent biocompatiblity and bioactivity and
thereby considered to be promising bone replacement material. ɛ-Polylysine (ɛ-PL) is non-toxic,
biodegradable, water soluble natural homopolymer with potential applications in medicine. In this
study, novel bioactive nanostructured composites for bone tissue engineering were developed based on
ɛ-PL and HAp. This composite biomaterial would combine the chemical and biological functionality of
ε-PL and bioactivity of HAp.
Nanostructured composites based on HAp and ɛ-PL were processed by spray drying technique using
slurry of in situ synthesized HAp in 5% (w/v) ɛ-PL aqueous solution. In situ synthesis of HAp was
realized by precipitation method using Ca2+ and PO43- ions sources as starting reagents with molar ratio
of Ca/P = 1.67 in order to obtain spray-dried products containing HAp and ɛ-PL at the 50/50, 70/30 and
90/10 wt%. The obtained slurry was spray dried by using a tabletop spray dryer with nozzle of 1.4 mm
diameter at a temperature ~190°C. The collected spray–dried composite powders were structurally
characterized using different analysis techniques.
SEM studies of obtained spray-dried products confirmed the formation of spherical microgranules with
diameter in the range 1-25 µm. Intimate interaction between HAp and ε-PL was evident from FT-IR and
SEM results. XRD patterns of fabricated composites revealed that nanocrystalline HAp was synthesized
in the presence of ε-PL. Accordingly, ε-PL influences HAp crystal nucleation and growth, allow for
chemical functionalization of the inorganic phase.
Novel composites based on HAp and ε-PL were fabricated using a combination of processing routes - in
situ precipitation and spray drying. Intimate interaction – adsorption of ε-polylysine on hydroxyapatite
crystallite surface is related with cationic nature of polymer.
Acknowledgements
This work has been supported by the National Research Programme No. 2014.10-4/VPP-3/21
“Multifunctional Materials and Composites, Photonics and Nanotechnology (IMIS2)” Project No. 4
44
THE ALLOCATION OF BONE FLUID IN BOVINE CORTICAL BONE
UTILIZING A MULTI-TECHNIQUE ANALYSIS
Luis Zelaya-Lainez1, Hawraa Kariem1, Winfried Nischkauer2, Andreas Limbeck2, Christian Hellmich1* 1Institute for Mechanics of Materials and Structures, Vienna University of Technology (TU Wien),
Karlsplatz 13/E202, 1040 Vienna, Austria. 2Institute of Chemical Technologies and Analytics, Division of Instrumental Analytical Chemistry,
Vienna University of Technology (TU Wien), Getreidemarkt 9/164, 1060 Vienna, Austria.
Abstract
The mechanical properties of bone depend crucially on the materials composition in terms of mineral
and collagen content, as well as the hierarchical organization of those components. What is less known,
is the role of the third major structural component in bone, which is bone fluid. It is partitioned in three
levels of pore spaces, ranging from the nanometer to the millimeter level. The porosity of cortical bone
includes the vascular porosity, the lacunae porosity, and the ultra-structure porosity. To determine the
partition of the major components, and specifically the fluid component, along the hierarchical structure
of bone, 24 bone samples were cut using a diamond band saw and low speed saw from a bovine femur.
The samples were polished in order to obtain images under a light microscope. The different porosities
were computed from the images. To calculate the major components in the samples, the bovine bone
samples were taken through a series of states, i.e. dehydrated state, rehydrated state with HBSS or
xylene, and demineralized state using a 0.5 M EDTA solution. The mass of the bone components are
normally given in terms of the content weight fraction, and for the samples rehydrated with HBSS the
mean weight fraction of the organic component is 0.21, of the mineral component 0.67, and of the fluid
component 0.12. The mean weight fraction of the fluid component within the vascular pores is 0.0161,
within the lacunae pores 0.0098, and within the ultra-structure pores 0.0870. For the samples rehydrated
with xylene, the mean weight fraction of the organic component is 0.20, of the mineral component 0.67,
and of the bone fluid component 0.13. The mean weight fraction of the fluid component within the
vascular pores is 0.02, within the lacunae pores 0.01, and within the ultra-structure pores 0.10. The
mean volume fraction for the samples rehydrated with HBSS given in term of the organic component is
0.33, of the mineral component 0.4510, and of the fluid component 0.22. The mean volume fraction for
the samples rehydrated with xylene given in term of the organic component is 0.29, of the mineral
component 0.42, and of the fluid component 0.29. The mean volume fraction of fluid within the
vascular pores is 0.03, within the lacunae pores 0.02, and within the ultra-structure pores 0.24.
45
Title of the conference – NEWGEN Zlín Workshop and WG Meeting On Hydrogel/ Biomineralized
Biomaterial for Bone Tissue Regeneration
Editor: Assoc. Prof. Nabanita Saha, PhD.
Published by Tomas Bata University in Zlin, 2016
1st Edition
ISBN:
